ArticleMicrobial cell factories2012
A strategy of gene overexpression based on tandem repetitive promoters in Escherichia coli.
Article in Microbial cell factories, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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40 citing papers in PubMed.
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- Analytical kinetic model of native tandem promoters in E. coli.PLoS computational biology · 2022Article
- Effects of Amino Acids and Overexpression of dapA Gene on the Production of ε-Poly-L-lysine by Streptomyces diastatochromogenes Strains.Current microbiology · 2021Article
- Poly(3-hydroxypropionate): Biosynthesis Pathways and Malonyl-CoA Biosensor Material Properties.Frontiers in bioengineering and biotechnology · 2021Review
- Characterization of Context-Dependent Effects on Synthetic Promoters.Frontiers in bioengineering and biotechnology · 2020Article
- Development of a novel strategy for robust synthetic bacterial promoters based on a stepwise evolution targeting the spacer region of the core promoter in Bacillus subtilis.Microbial cell factories · 2019Article
- High-level extracellular production of recombinant nattokinase in Bacillus subtilis WB800 by multiple tandem promoters.BMC microbiology · 2019Article
- T5 exonuclease-dependent assembly offers a low-cost method for efficient cloning and site-directed mutagenesis.Nucleic acids research · 2019Article
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7 authors.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundFor metabolic engineering, many rate-limiting steps may exist in the pathways of accumulating the target metabolites. Increasing copy number of the desired genes in these pathways is a general method to solve the problem, for example, the employment of the multi-copy plasmid-based expression system. However, this method may bring genetic instability, structural instability and metabolic burden to the host, while integrating of the desired gene into the chromosome may cause inadequate transcription or expression. In this study, we developed a strategy for obtaining gene overexpression by engineering promoter clusters consisted of multiple core-tac-promoters (MCPtacs) in tandem.
resultsThrough a uniquely designed in vitro assembling process, a series of promoter clusters were constructed. The transcription strength of these promoter clusters showed a stepwise enhancement with the increase of tandem repeats number until it reached the critical value of five. Application of the MCPtacs promoter clusters in polyhydroxybutyrate (PHB) production proved that it was efficient. Integration of the phaCAB genes with the 5CPtacs promoter cluster resulted in an engineered E.coli that can accumulate 23.7% PHB of the cell dry weight in batch cultivation.
conclusionsThe transcription strength of the MCPtacs promoter cluster can be greatly improved by increasing the tandem repeats number of the core-tac-promoter. By integrating the desired gene together with the MCPtacs promoter cluster into the chromosome of E. coli, we can achieve high and stale overexpression with only a small size. This strategy has an application potential in many fields and can be extended to other bacteria.
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